The HORIBA Scientific SPRi instruments are ideal solutions for label-free and multiplexed biomolecular analysis. These compact and flexible devices capable of either automated or manual operation are designed for easy determination of real-time interaction and kinetic studies. Their open formats enable numerous types of experiments covering biomolecular interactions, biochemistry, chemistry and physical-chemistry, to be explored without restriction or compromise.
This application note shows how binding kinetics of single-domain antibodies can be studied with SPRi technology. QVQ produces single-domain antibodies from camelids (sdAbs or VHH). Like conventional Abs, sdAbs are able to bind specific epitopes with high binding affinity. The small size of sdAbs allows for enhanced tumor penetration and fast blood clearance, features that are favorable for in vivo imaging applications. Here, the binding of the anti-HER2 sdAb Q17c to recombinant HER2 protein was assessed using Surface Plasmon Resonance imaging (SPRi). An optimization of immobilization conditions was performed for Q17c and its specific antigen HER2 using a single SPRi-Biochip.
A monoclonal antibody (mAb) highly specific to a steroid hormone (undisclosed) of 290 Daltons was analyzed thanks to SPRi technology. This application note highlights the sensitivity of the XelPleXTM system for the detection of small molecules.
Surface plasmon resonance imaging allows molecules binding study in label-free and real-time conditions. The power of the technique comes from its singularities, which are multiplexing and imaging, leading to a considerable speed-up of the processes’ analyses. This will be illustrated for antibody molecule study by three applications.
This article has been published in the Spring issue of the International Biopharmaceutical Industry (IBI) journal.
DARPins (Designed Ankyrin Repeat Proteins) are a class of non-immunoglobulin binders. Thanks to their specificity and robustness, they allow a multitude of novel, so far unfeasible applications. Surface Plasmon Resonance imaging (SPRi) is a powerful label-free technique that enables real-time target detection. Combining SPRi to massspectrometry (MS) allows biomolecules identification using their unique peptide mass fingerprint. In the past this combination was cumbersome and time-consuming. This application note shows how a protein kinase (RPS6KA2), a potential drug target is detected and identified using a hyphenated On-Chip SPR-MS coupling protocol, leading to saving time and money.